Loop-mediated isothermal amplification in detection of West Nile virus genome / 第二军医大学学报
Academic Journal of Second Military Medical University
;
(12): 590-594, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-840279
ABSTRACT
Objective:
To establish a loop-mediated isothermal amplification (LAMP) assay for rapid detection of the West Nile virus (WNV).Methods:
WNV genome (position nt 1 021 to nt 1 240) was synthesized by a PCR-based gene synthesis method. The synthetic fragments included 6 pairs of LAMP primer recognizing 8 primer sites of WNV genome. The LAMP gene amplification was carried out using a real-time PCR system at 63°C for 60 min, then the amplification was terminated at 80°C after 2 min. The amplification products were observed by agarose gel electrophoresis. The sensitivity and specificity of LAMP assay were compared with those of conventional PCR.Results:
The LAMP assay took less than 20 min, and the amplification product took on a ladder-like electrophoresis pattern. The sensitivity of LAMP assay was 10-fold higher than that of conventional PCR, and the detection limit of LAMP was 9.23 copies/μl. The specificity of WNV-specific LAMP assay was demonstrated by the negative amplification results from dengue virus and Japanese encephalitis virus, both were closely related members of the Flavivirus family.Conclusion:
LAMP assay is rapid, cost-effective, highly sensitive and specific in detecting genes of interest, and is of great significance for WNV surveillance, especially for grass root units and on-sport surveillance.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Tipo de estudo:
Estudo diagnóstico
Idioma:
Chinês
Revista:
Academic Journal of Second Military Medical University
Ano de publicação:
2010
Tipo de documento:
Artigo
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